Method of treating primary sclerosing cholangitis

ABSTRACT

Disclosed is a method for treating a subject having primary sclerosing cholangitis, comprising administering to said subject an effective amount of a humanized antibody or antigen-binding fragment thereof having binding specificity for alpha 4 beta 7 integrin.

RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No.62/129,698, filed on Mar. 6, 2015. The entire contents of the foregoingapplication are hereby incorporated by reference.

BACKGROUND

Vedolizumab and other agents that bind α4 integrin, as well as TNFinhibitors, have been used to treat inflammatory bowel diseases, butthese agents have not been used to treat sclerotic diseases of theliver, such as primary sclerosing cholangitis (PSC).

PSC is a chronic cholestatic hepatobiliary disease characterized byinflammation and progressive fibrosis of intra- and extra-hepatic bileducts leading to biliary cirrhosis, portal hypertension, and finallyliver failure. As with many chronic liver diseases, PSC progressesinsidiously even in asymptomatic patients. There is currently noeffective treatment and therapy options are limited to managingcomplications: e.g., dilatation of dominant strictures and earlydiagnosis of cholangiocarcinoma. Orthotopic liver transplantation iscurrently the only life-extending therapy for patients progressing toliver failure.

PSC is a disease of relatively young people, with a median age atdiagnosis of 40 years. PSC has a profound impact on patients and isaccompanied by substantial longstanding morbidity, before livertransplantation or death ensues. Overall PSC is relatively uncommon(annual incidence of 1:100,000), however, the natural history of thedisease and mean survival rates (˜12 years from diagnosis) ensure thatdisease prevalence approaches 8-14 in 100,000.

PSC has an unknown etiology, however one hypothesis suggests that it isimmune- mediated, resulting in an erroneous chronic inflammatoryresponse against the biliary epithelium. About 70% to 80% of PSCpatients also suffer from inflammatory bowel disease (IBD), such asulcerative colitis (UC), Crohn's disease (CD), and IBD Unclassified(IBDU). Patients suffering from both PSC and IBD are viewed as distinctfrom a number of other minor PSC subtypes, e.g., immunoglobulin G4(IgG4)-positive sclerosing cholangitis; PSC with autoimmune hepatitis;small-duct associated PSC.

PSC represents a disease of very high unmet medical need, which despitenumerous studies with various agents (Karlsen et al., Aliment.Pharmacol. Ther. 39:282-301 (2014)), has no therapy shown to preventdisease progression. Treatment options are limited to attempting tomanage ongoing symptoms and complications. This places patients with PSCat risk from liver failure and ultimately death, with only livertransplantation as a potential option. Thus, a need exists for improvedtherapeutic approaches to primary sclerosing cholangitis.

SUMMARY

In a first aspect, the invention relates to a method for treating ahuman subject suffering from primary sclerosing cholangitis (PSC),comprising the steps of: administering to a subject suffering from PSC,an effective amount of an anti-α4β7 antibody, wherein the anti-α4β7antibody has binding specificity for the α4β7 complex, and wherein theantigen-binding region comprises the CDRs: light chain: CDR1 SEQ IDNO:11, CDR2 SEQ ID NO:12, CDR3 SEQ ID NO:13; heavy chain: CDR1 SEQ IDNO:8, CDR2 SEQ ID NO:9, and CDR3 SEQ ID NO:10.

In some embodiments, an effective amount is an amount sufficient tonormalize serum alkaline phosphatase (ALP) of the subject. In someembodiments, an effective amount is an amount sufficient to reduce serumalkaline phosphatase (ALP) of the subject by at least 35%. In someembodiments, an effective amount is an amount sufficient to improveIshak necroinflammatory grading score by at least one point.

In some embodiments, the anti-α4β7 antibody may be administered to thesubject according to the following dosing regimen: (a) an initial doseof 300 mg of the anti-α4β7 antibody as an intravenous infusion; (b)followed by a second subsequent dose of 300 mg of the anti-α4β7 antibodyas an intravenous infusion at about two weeks after the initial dose;(c) followed by a third subsequent dose of 300 mg of the anti-α4β7antibody as an intravenous infusion at about six weeks after the initialdose; (d) followed by a fourth and subsequent doses of 300 mg of theanti-α4β7 antibody as an intravenous infusion every four weeks or everyeight weeks after the third subsequent dose of the anti-α4β7 antibody asneeded.

In some embodiments, the subject has chronic cholestatic liver diseasewith a subsequent diagnosis of PSC based on cholangiographic findings ofintrahepatic and/or extrahepatic bile duct irregularities consistentwith PSC.

In some embodiments, the subject further has diagnosis of IBD byclinical and endoscopic evidence and corroborated by a histopathologyreport. In some embodiments, the subject does not have a diagnosis ofIBD.

In some embodiments, the subject has alkaline phosphatase (ALP)elevation to at least 1.6 times the upper limit of normal (ULN) at thetime of initial treatment.

In some embodiments, the subject's Ishak fibrosis staging score isimproved, maintained or normalized.

In some embodiments, the subject's Amsterdam Cholestatic Complaints(ACCS) has improved, maintained or normalized, 5-D itch scale hasimproved, maintained or normalized, or the subject has a liver stiffnessTE score of less than or equal to 14.3 kPa, as assessed by VibrationControlled Transient Elastrography.

In some embodiments, the anti-α4β7 antibody is administered to thesubject intravenously. In other embodiments, the anti-α4β7 antibody isadministered subcutaneously. In some embodiments, the anti-α4β7 antibodyis administered intravenously during part of the therapeutic regimen andsubcutaneously during part of the therapeutic regimen.

In some embodiments, a PSC-related outcome selected from the groupconsisting of progression to cirrhosis, liver failure, death and livertransplantation is delayed or prevented. In some embodiments, aPSC-related complication selected from the group consisting of ascites,hepatic encephalopathy, development of varices, jaundice, varicealbleeding, cholangiocarcinoma, hepatocellular carcinoma, evidence ofcirrhosis, and colorectal cancer is delayed or prevented.

In some embodiments, the treatment does not cause one or more than oneadverse event selected from the group consisting of hepatoxicity, PML,cholangiocarcinoma, one or more complications due to portalhypertension, leucopenia, lymphopenia, colorectal cancer,infusion-related reactions, infection, acute respiratory failure, acuterespiratory distress syndrome, Torsade de pointes, ventricularfibrillation, ventricular tachycardia, malignant hypertension,convulsive seizure, agranulocytosis, aplastic anemia, toxic epidermalnecrolysis, Stevens-Johnson syndrome, hepatic necrosis, acute liverfailure, anaphylactic shock, acute renal failure, pulmonaryhypertension, pulmonary fibrosis, confirmed or suspected endotoxinshock, confirmed or suspected transmission of infectious agent by amedicinal product, neuroleptic malignant syndrome, malignanthyperthermia, spontaneous abortion, stillbirth, and fetal death.

In some embodiments, the anti-α4β7 antibody has a heavy chain variableregion sequence comprising amino acid residues 20 to 140 of SEQ ID NO:2,and a light chain variable region comprising amino acids 20 to 131 ofSEQ ID NO:4 or amino acids 21 to 132 of SEQ ID NO:5. In someembodiments, the anti-α4β7 antibody is vedolizumab.

In another aspect, the invention relates to a pharmaceutical compositioncomprising an antibody having binding specificity for human α4β7integrin for the manufacture of a medicament for treating primarysclerosing cholangitis.

In another aspect, the invention relates to a pharmaceutical compositioncomprising an antibody having binding specificity for human α4β7integrin for use in treating primary sclerosing cholangitis.

In some embodiments, the pharmaceutical composition may be formulated asa lyophilized preparation. In some embodiments, the pharmaceutical maybe formulated as a liquid preparation.

In some embodiments, the pharmaceutical composition may be formulated ina device for subcutaneous administration.

In another aspect, the invention relates to a therapeutically effectiveamount of an anti-α4β7 antibody for use in the treatment or prophylaxisof PSC in an individual in need thereof.

In another aspect, the invention relates to use of a therapeuticallyeffective amount of an anti-α4β7 antibody for the manufacture of amedicament for use in the treatment or prophylaxis of PSC in a subjectin need thereof.

In some embodiments, the anti-α4β7 antibody is humanized.

DETAILED DESCRIPTION

The invention relates to methods of treating primary sclerosingcholangitis (PSC) by administering an anti-α4β7 antibody. PSC isdistinguished from secondary sclerosing cholangitis (SSC). Patientssuffering from PSC at first can be asymptomatic, with no physicalabnormalities, but over time, death results from cancer or liverfailure, unless the patient undergoes liver transplantation. Patientssuffering from PSC additionally can be suffering from IBD. “PSC withunderlying IBD” is a term which indicates a diagnosis comprising bothconditions, but does not indicate IBD as the cause of PSC or IBD as anecessary association with PSC. The methods and compositions related totreatment with an anti-α4β7 antibody can improve the outcome for PSCpatients.

Definitions

The term “pharmaceutical formulation” refers to a preparation thatcontains an anti-α4β7 antibody in such form as to permit the biologicalactivity of the antibody to be effective, and which contains noadditional components which are unacceptably toxic to a subject to whichthe formulation would be administered.

The cell surface molecule, “α4β7 integrin,” or “α4β7,” is a heterodimerof an α₄ chain (CD49D, ITGA4) and a β₇ chain (ITGB7). Each chain canform a heterodimer with an alternative integrin chain, to form α₄β₁ orα_(E)β₇. Human α₄ and β₇ genes (GenBank (National Center forBiotechnology Information, Bethesda, Md.) RefSeq Accession numbersNM_000885 and NM_000889, respectively) are expressed by B and Tlymphocytes, particularly memory CD4+ lymphocytes. Typical of manyintegrins, α4β7 can exist in either a resting or activated state.Ligands for α4β7 include vascular cell adhesion molecule (VCAM),fibronectin and mucosal addressin (MAdCAM (e.g., MAdCAM-1)).

As used herein, a human antibody or antigen-binding fragment thereofthat has “binding specificity for the α4β7 complex” binds to α4β7, butnot to α4β1 or αEβ7.

The term “antibody” herein is used in the broadest sense andspecifically covers full length monoclonal antibodies, immunoglobulins,polyclonal antibodies, multispecific antibodies (e.g. bispecificantibodies) formed from at least two full length antibodies, e.g., eachto a different antigen or epitope, and individual antigen bindingfragments, including dAbs, scFv, Fab, F(ab)′₂, Fab′, including human,humanized and antibodies from non-human species and recombinant antigenbinding forms such as monobodies and diabodies.

The term “human antibody” includes an antibody that possesses a sequencethat is derived from a human germ-line immunoglobulin sequence, such asan antibody derived from transgenic mice having human immunoglobulingenes (e.g., XENOMOUSE genetically engineered mice (Abgenix, Fremont,Calif.), HUMAB-MOUSE®, KIRIN TC MOUSE™ transchromosome mice, KMMOUSE®(MEDAREX, Princeton, N.J.)), human phage display libraries, humanmyeloma cells, or human B cells.

The term “monoclonal antibody” as used herein refers to an antibodyobtained from a population of substantially homogeneous antibodies,i.e., the individual antibodies comprising the population are identicaland/or bind the same epitope, except for possible variants that mayarise during production of the monoclonal antibody, such variantsgenerally being present in minor amounts. In contrast to polyclonalantibody preparations that typically include different antibodiesdirected against different determinants (epitopes), each monoclonalantibody is directed against a single determinant on the antigen. Themodifier “monoclonal” indicates the character of the antibody as beingobtained from a substantially homogeneous population of antibodies, andis not to be construed as requiring production of the antibody by anyparticular method. For example, the monoclonal antibodies to be used inaccordance with the present invention may be made by the hybridomamethod first described by Kohler et al., Nature, 256:495 (1975), or maybe made by recombinant DNA methods (see, e.g., U.S. Pat. No. 4,816,567).The “monoclonal antibodies” may also be isolated from phage antibodylibraries using the techniques described in Clackson et al., Nature,352:624-628 (1991) and Marks et al., J. Mol. Biol., 222:581-597 (1991),for example.

The monoclonal antibodies herein specifically include “chimeric”antibodies in which a portion of the heavy and/or light chain isidentical with or homologous to corresponding sequences in antibodiesderived from a particular species or belonging to a particular antibodyclass or subclass, while the remainder of the chain(s) is identical withor homologous to corresponding sequences in antibodies derived fromanother species or belonging to another antibody class or subclass, aswell as fragments of such antibodies, so long as they exhibit thedesired biological activity (U.S. Pat. No. 4,816,567; and Morrison etal., Proc. Natl. Acad. Sci. USA, 81:6851-6855 (1984)). Chimericantibodies of interest herein include “primatized” antibodies comprisingvariable domain antigen binding sequences derived from a non-humanprimate (e.g. Old World Monkey, Ape etc) and human constant regionsequences.

“Antigen binding fragments” of the e.g., humanized, antibody prepared inthe formulation, use and/or method of the invention comprise at leastthe variable regions of the heavy and/or light chains of an anti-α4β7antibody. For example, an antigen binding fragment of vedolizumabcomprises amino acid residues 20-131 of the humanized light chainsequence of SEQ ID NO:4. Examples of such antigen binding fragmentsinclude Fab fragments, Fab′ fragments, scFv and F(ab′)₂ fragments of anantibody known in the art. Antigen binding fragments of the anti-α4β7antibody of the invention can be produced by enzymatic cleavage or byrecombinant techniques. For instance, papain or pepsin cleavage can beused to generate Fab or F(ab′)₂ fragments, respectively. Antibodies canalso be produced in a variety of truncated forms using antibody genes inwhich one or more stop codons have been introduced upstream of thenatural stop site. For example, a recombinant construct encoding theheavy chain of an F(ab′)₂ fragment can be designed to include DNAsequences encoding the CH₁ domain and hinge region of the heavy chain.In one aspect, antigen binding fragments inhibit binding of α4β7integrin to one or more of its ligands (e.g. the mucosal addressinMAdCAM (e.g., MAdCAM-1), fibronectin).

Papain digestion of antibodies produces two identical antigen bindingfragments, called “Fab” fragments, each with a single antigen bindingsite, and a residual “Fc” fragment, whose name reflects its ability tocrystallize readily. Pepsin treatment yields an F(ab′)₂ fragment thathas two antigen binding sites and is still capable of cross-linkingantigen.

“Fv” is an antibody fragment which consists of a dimer of one heavychain variable domain and one light chain variable domain innon-covalent association.

The Fab fragment also contains the constant domain of the light chainand the first constant domain (CH1) of the heavy chain. Fab′ fragmentsdiffer from Fab fragments by the addition of a few residues at thecarboxy terminus of the heavy chain CH1 domain including one or morecysteines from the antibody hinge region. Fab′-SH is the designationherein for Fab′ in which the cysteine residue(s) of the constant domainsbear at least one free thiol group. F(ab′)₂ antibody fragmentsoriginally were produced as pairs of Fab′ fragments which have hingecysteines between them. Other chemical couplings of antibody fragmentsare also known.

“Single-chain Fv” or “scFv” antibody fragments comprise the V_(H) andV_(L) domains of antibody, wherein these domains are present in a singlepolypeptide chain. In one aspect, the Fv polypeptide further comprises apolypeptide linker between the V_(H) and V_(L) domains which enables thescFv to form the desired structure for antigen binding. For a review ofscFv see Pluckthun in The Pharmacology of Monoclonal Antibodies, vol.113, Rosenburg and Moore eds., Springer-Verlag, New York, pp. 269-315(1994).

The term “diabodies” refers to small antibody fragments with two antigenbinding sites, which fragments comprise a variable heavy domain (V_(H))connected to a variable light domain (V_(L)) in the same polypeptidechain (V_(H)-V_(L)). By using a linker that is too short to allowpairing between the two domains on the same chain, the domains areforced to pair with the complementary domains of another chain andcreate two antigen binding sites. Diabodies are described more fully in,for example, EP 404,097; WO 93/11161; and Hollinger et al., Proc. Natl.Acad. Sci. USA, 90:6444-6448 (1993).

A “full length antibody” is one which comprises an antigen bindingvariable region as well as a light chain constant domain (C_(L)) andheavy chain constant domains, C_(H1), C_(H2) and C_(H3). The constantdomains may be native sequence constant domains (e.g. human nativesequence constant domains) or amino acid sequence variants thereof. Inone aspect, the full length antibody has one or more effector functions.

An “amino acid sequence variant” antibody herein is an antibody with anamino acid sequence which differs from a main species antibody.Ordinarily, amino acid sequence variants will possess at least about70%, at least about 80%, at least about 85%, at least about 90%, or atleast about 95% homology with the main species antibody. The amino acidsequence variants possess substitutions, deletions, and/or additions atcertain positions within or adjacent to the amino acid sequence of themain species antibody, but retain antigen binding activity. Variationsin sequence of the constant regions of the antibody will have lesseffect on the antigen binding activity than variations in the variableregions. In the variable regions, amino acid sequence variants will beat least about 90% homologous, at least about 95% homologous, at leastabout 97% homologous, at least about 98% homologous, or at least about99% homologous with the main species antibody.

“Homology” is defined as the percentage of residues in the amino acidsequence variant that are identical after aligning the sequences andintroducing gaps, if necessary, to achieve the maximum percent homology.Methods and computer programs for the alignment are well known in theart.

A “therapeutic monoclonal antibody” is an antibody used for therapy of ahuman subject. Therapeutic monoclonal antibodies disclosed hereininclude anti-α4β7 antibodies.

Depending on the amino acid sequence of the constant domain of theirheavy chains, full length antibodies can be assigned to different“classes”. There are five major classes of full length antibodies: IgA,IgD, IgE, IgG, and IgM, and several of these may be further divided into“subclasses” (isotypes), e.g., IgG1, IgG2, IgG3, IgG4, IgA, and IgA2.The heavy-chain constant domains that correspond to the differentclasses of antibodies are called α, δ, ε, γ, and μ, respectively. Thesubunit structures and three-dimensional configurations of differentclasses of immunoglobulins are well known.

The “light chains” of antibodies from any vertebrate species can beassigned to one of two clearly distinct types, called kappa (κ) andlambda (λ), based on the amino acid sequences of their constant domains.

The terms “Fc receptor” or “FcR” are used to describe a receptor thatbinds to the Fc region of an antibody. In one aspect, the FcR is anative sequence human FcR. In another aspect, the FcR is one which bindsan IgG antibody (a gamma receptor) and includes receptors of the FcγRI,FcγRII, and FcγRIII subclasses, including allelic variants andalternatively spliced forms of these receptors. FcγRII receptors includeFcγRIIA (an “activating receptor”) and FcγRIIB (an “inhibitingreceptor”), which have similar amino acid sequences that differprimarily in the cytoplasmic domains thereof. Activating receptorFcγRIIA contains an immunoreceptor tyrosine-based activation motif(ITAM) in its cytoplasmic domain. Inhibiting receptor FcγRIIB containsan immunoreceptor tyrosine-based inhibition motif (ITIM) in itscytoplasmic domain. (See review in M. Daeron, Annu. Rev. Immunol.15:203-234 (1997)). FcRs are reviewed in Ravetch and Kinet, Annu. Rev.Immunol 9:457-92 (1991); Capel et al., Immunomethods 4:25-34 (1994); andde Haas et al., J. Lab. Clin. Med. 126:33-41 (1995). Other FcRs,including those to be identified in the future, are encompassed by theterm “FcR” herein. The term also includes the neonatal receptor, FcRn,which is responsible for the transfer of maternal IgGs to the fetus(Guyer et al., J. Immunol. 117:587 (1976) and Kim et al., J. Immunol.24:249 (1994)) and for regulating the persistence of immunoglobulin G(IgG) and albumin in the serum (reviewed by Rath et al., J. Clin.Immunol. 33 Suppl 1:S9-17 (2013)).

The term “hypervariable region” when used herein refers to the aminoacid residues of an antibody which are responsible for antigen bindingand are found in the “variable domain” of each chain. The hypervariableregion generally comprises amino acid residues from a “complementaritydetermining region” or “CDR” (e.g. residues 24-34 (L1), 50-56 (L2) and89-97 (L3) in the light chain variable domain and 31-35 (H1), 50-65 (H2)and 95-102 (H3) in the heavy chain variable domain; Kabat et al.,Sequences of Proteins of Immunological Interest, 5th Ed. Public HealthService, National Institutes of Health, Bethesda, Md. (1991)) and/orthose residues from a “hypervariable loop” (e.g. residues 26-32 (L1),50-52 (L2) and 91-96 (L3) in the light chain variable domain and 26-32(H1), 53-55 (H2) and 96-101 (H3) in the heavy chain variable domain;Chothia and Lesk J. Mol. Biol. 196:901-917 (1987)). “Framework Region”or “FR” residues are those variable domain residues other than thehypervariable region residues as herein defined. The hypervariableregion or the CDRs thereof can be transferred from one antibody chain toanother or to another protein to confer antigen binding specificity tothe resulting (composite) antibody or binding protein.

“Humanized” forms of non-human (e.g., rodent) antibodies are chimericantibodies that contain minimal sequence derived from non-humanimmunoglobulin. For the most part, humanized antibodies are humanimmunoglobulins (recipient antibody) in which residues from ahypervariable region of the recipient are replaced by residues from ahypervariable region of a non-human species (donor antibody) such asmouse, rat, rabbit or nonhuman primate having the desired specificity,affinity, and capacity. In some instances, framework region (FR)residues of the human immunoglobulin are replaced by correspondingnon-human residues. Furthermore, humanized antibodies may compriseresidues that are not found in the recipient antibody or in the donorantibody. These modifications are made to further refine antibodyperformance. In general, the humanized antibody will comprisesubstantially all of at least one, and typically two, variable domains,in which all or substantially all of the hypervariable loops correspondto those of a non-human immunoglobulin and all or substantially all ofthe FRs are those of a human immunoglobulin sequence. The humanizedantibody optionally also will comprise at least a portion of animmunoglobulin constant region (Fc), typically that of a humanimmunoglobulin. For further details, see Jones et al., Nature321:522-525 (1986); Riechmann et al., Nature 332:323-329 (1988); andPresta, Curr. Op. Struct. Biol. 2:593-596 (1992).

An “affinity matured” antibody is one with one or more alterations inone or more hypervariable regions thereof which result an improvement inthe affinity of the antibody for antigen, compared to a parent antibodywhich does not possess those alteration(s). In one aspect, affinitymatured antibodies will have nanomolar or even picomolar affinities forthe target antigen. Affinity matured antibodies are produced byprocedures known in the art. Marks et al. Bio/Technology 10:779-783(1992) describes affinity maturation by VH and VL domain shuffling.Random mutagenesis of CDR and/or framework residues is described by:Barbas et al. Proc Nat. Acad. Sci, USA 91:3809-3813 (1994); Schier etal. Gene 169:147-155 (1995); Yelton et al. J. Immunol. 155:1994-2004(1995); Jackson et al., J. Immunol. 154(7):3310-9 (1995); and Hawkins etal., J. Mol. Biol. 226:889-896 (1992).

An “isolated” antibody is one which has been identified and separatedand/or recovered from a component of its natural environment. In certainembodiments, the antibody will be purified (1) to greater than 95% byweight of protein as determined by the Lowry method, and alternatively,more than 99% by weight, (2) to a degree sufficient to obtain at least15 residues of N-terminal or internal amino acid sequence by use of aspinning cup sequenator, or (3) to homogeneity by SDS-PAGE underreducing or nonreducing conditions using Coomassie blue or silver stain.Isolated antibody includes the antibody in situ within recombinant cellssince at least one component of the antibody's natural environment willnot be present. Ordinarily, however, isolated antibody will be preparedby at least one purification step.

“Treatment” refers to both therapeutic treatment and prophylactic orpreventative measures. Treatment refers to any type of measure thatimparts a benefit to a subject or a patient afflicted with a disease,including improvement in the condition of the subject or patient (e.g.,in one or more symptoms). Treatment may include any drug, drug product,method, procedure, lifestyle change, or other adjustment introduced inan attempt to effect a change in a particular aspect of a subject'shealth (i.e., directed to a particular disease, disorder, or condition).Those in need of treatment include those already with the disease aswell as those in which the disease or its recurrence is to be prevented.Hence, the patient to be treated herein may have been diagnosed ashaving the disease or may be predisposed or susceptible to the disease.The terms “patient,” “individual” and “subject” are used interchangeablyherein.

As used herein, the term “a therapeutically effective amount” means anamount of the composition comprising the therapeutic agent that willaffect a treatment of the subject as defined above. In some embodiments,a therapeutically effective amount improves the condition or delays theonset or progress of the condition of a subject without causing asignificant side-effect or adverse event.

As used herein, “a healthy subject” means a subject that has not beendiagnosed with PSC and/or does not exhibit any detectable symptomsassociated with PSC.

The antibody which is formulated is substantially pure and desirablysubstantially homogeneous (i.e. free from contaminating proteins etc).“Substantially pure” antibody means a composition comprising at leastabout 90% antibody by weight, based on total weight of the protein inthe composition, at least about 95% or 97% by weight. “Substantiallyhomogeneous” antibody means a composition comprising protein wherein atleast about 99% by weight of protein is specific antibody, e.g.,anti-α4β7 antibody, based on total weight of the protein.

Treatment of PSC with Anti-α4β7 Antibodies

In one aspect, the invention relates to a method of treating PSC in asubject comprising administering to the subject an anti-α4β7 antibodydescribed herein in an amount effective to treat PSC, e.g., in humans.The human subject may be an adult (e.g., 18 years or older), anadolescent, or a child. A patient suffering from PSC who can benefitfrom anti-α4β7 integrin antibody therapy can have abnormal liverfunction tests. For example, the patient can have an abnormal alkalinephosphatase test. In a PSC patient who can benefit from anti-α4β7integrin antibody therapy, the alkaline phosphatase level can be greaterthan the upper limit of normal (ULN), e.g. 1.5 times ULN, 1.6 times ULN,2 times ULN, 2.5 times ULN, 3 times ULN, 4 times ULN, or a range of 1.5to 10 times ULN or a range of 3 to 12 times ULN. Other abnormal liverfunction tests which can be exhibited by a patient suffering from PSCinclude a test selected from the group consisting of alaninetransaminase, γ-Glutamyl transpeptidase, aspartate transaminase, andtotal bilirubin. In some embodiments, the method relates to treating aPSC patient with an anti-α4β7 integrin antibody if the patient has anabnormal liver function test. In some embodiments, the method relates totreating a PSC patient with an anti-α4β7 integrin antibody if thepatient has an abnormal alkaline phosphatase test. In some embodiments,the patient has elevated ALP that is not attributable to a hepatic ornonhepatic cause. In a further embodiment, the PSC patient can haveliver fibrosis or IBD. The IBD can be ulcerative colitis, Crohn'sdisease or indeterminate, undifferentiated or unclassified IBD (IBDU). Apatient suffering from PSC who can benefit from anti-α4β7 integrinantibody therapy can have abnormal liver stiffness. In some embodiments,the method relates to treating a PSC patient with an anti-α4β7 integrinantibody if the patient has a liver stiffness TE score of ≦20 kPa, ≦18kPa, ≦16 kPa, ≦15 kPa, ≦14 kPa, ≦13 kPa. In an embodiment, a PSC patientis treated with an anti-α4β7 antibody if the liver stiffness TE score is≦14.3 kPa. In some embodiments, the patient does not have elevated serumIgG4, e.g., at least 2 times its ULN or the IgG4/IgG1 ratio is above0.24.

A pharmaceutical composition comprising an anti-α4β7 antibody can beused as described herein for treating PSC in a subject sufferingtherefrom. Typical PSC related clinical outcomes include, for example,progression to cirrhosis, liver failure, death and livertransplantation. PSC related clinical complications include, forexample, ascites, hepatic encephalopathy, development of varices,jaundice, variceal bleeding, cholangiocarcinoma, hepatocellularcarcinoma, evidence of cirrhosis, and colorectal cancer. The method andpharmaceutical compositions described herein can improve clinicaloutcome and/or clinical complications of PSC.

An effective amount of an anti-α4β7 antibody of the present invention isadministered to an individual, e.g. a subject, in need thereof (e.g., amammal, such as a human or other primate) in order to treat PSC. Aneffective amount of an anti-α4β7 antibody may be an amount sufficient toreduce, delay or prevent progression of PSC-related clinicalcomplications, liver failure and/or death. For example, the effectiveamount of anti-α4β7 antibody can be an amount that is sufficient tomaintain (e.g., prevent worsening), improve or normalize a clinicaldisease assessment score, or to maintain, reduce or normalize the levelof a marker of liver function and/or pathology in the individual. Insome aspects, the amount of anti-α4β7 antibody that is administered issufficient to maintain, improve an Ishak fibrosis staging score, anamount sufficient to maintain, reduce or normalize serum alkalinephosphatase (ALP), an amount sufficient to maintain, improve Ishaknecroinflammatory grading score, an amount sufficient to maintain,improve or normalize Amsterdam Cholestatic Complaints Score (ACCS), anamount sufficient to maintain, improve or normalize 5-D itch scale, anamount sufficient to maintain, improve or normalize the time toprogression to cirrhosis by transient elastography (TE) assessed usingFibroScan®, an amount sufficient to maintain, improve or normalize thetime to PSC-related clinical outcomes or clinical complications, anamount sufficient to maintain, improve or normalize a subject's collagenproportional area (CPA, e.g., as assessed by histology), an amountsufficient to maintain, improve or normalize Enhanced Liver Fibrosis(ELF) score (e.g., as assessed by an algorithm using tests for serumconcentrations of procollagen-III aminoterminal propeptide, tissueinhibitor of matrix metalloproteinase-1 and hyaluronic acid), an amountsufficient to maintain, improve or normalize liver stiffness score byTransient Elastography, an amount sufficient to maintain, improve ornormalize liver stiffness score by Magnetic Resonance Elastography(MRE), an amount sufficient to maintain, improve or normalize Mayo PSCrisk score, or any combination thereof.

Certain markers can be used as metrics to administer an effective amountof anti-α4β7 antibody. For example, in some aspects an amount ofanti-α4β7 antibody that is sufficient to maintain, reduce or normalize asubject's serum alanine aminotransferase (ALT), aspartateaminotransferase (AST), gamma-glutamyl transpeptidase (GGT), bilirubin,vascular adhesion protein-1 (VAP-1), or any combination thereof, isadministered. Alternatively or in addition, an effective amount ofanti-α4β7 antibody may be an amount sufficient to maintain, improve ornormalize a subject's platelet ratio index (APRI) or Fibrosis 4 (FIB-4)score. In one embodiment, the invention relates to a method of treatingPSC in a subject comprising administering to a subject an anti-α4β7antibody described herein in an amount effective to reduce serumalkaline phosphatase (ALP) by at least about 20%, at least about 25%, atleast about 30%, at least about 35%, at least about 40%, or at leastabout 45%. An effective amount of anti-α4β7 antibody may be an amountsufficient to normalize serum ALP in a subject. An effective amount ofanti-α4β7 antibody may be an amount sufficient to reduce serum ALP in asubject to less than the Upper Limit of Normal (ULN). Reference range inhuman adults is 40 - 125 IU/L (VITROS® AnalyzerP, Ortho ClinicalDiagnostics, Rochester, N.Y.) or about 40 to about 150 IU/L (othermethods). Less than ULN would be less than 200 IU/L, less than 175 IU/L,less than 150 IU/L, less than 145 IU/L, less than 140 IU/L, less than135 IU/L, less than 130 IU/L or less than 125 IU/L.

In one embodiment, the invention relates to a method of treating PSC ina subject comprising administering to a subject an anti-α4β7 antibodydescribed herein in an amount effective to improve the subject's Ishaknecroinflammatory grading score by at least 1 or more points (e.g, 1, 2,3, 4 points). In another embodiment, the invention relates to a methodof treating PSC in a subject comprising administering to a subject ananti-α4β7 antibody described herein in an amount effective to preventworsening of the subject's Ishak necroinflammatory grading score.

In other embodiments, an effective amount of anti-α4β7 antibody may bean amount sufficient to improve the 5-D itch score or the AmsterdamCholestatic Complaints Score (ACCS) in a subject.

An endpoint such as no worsening of fibrosis, utilizes histology withIshak fibrosis staging as a measure of structural and architecturalalterations. PSC studies, both retrospective and prospective indicatethat about ˜50% of PSC patients will experience progression of fibrosisover 2 years. To date histology, via a liver biopsy, has been used toassess the progression of various liver diseases and has been includedin clinical trials assessing the efficacy of new therapeutic agents. Thescoring system proposed by Ishak has gained the most attention as itincorporates concepts relating to the pathogenesis of liver damage(inflammation and fibrosis), provides a wide range of possibleassessment scores, and is also one of the few histology scoring systemsto be shown to have prognostic value to long-term clinical outcomes.

Reductions or normalization of ALP levels have been demonstrated as aprognosis factor in PSC, being associated with increased overallsurvival times, increased liver transplant-free survival, and alsoreduced risk of cholangiocarcinoma.

Liver stiffness can be indicative of the extent of fibrosis in theliver. Healthy liver is soft or elastic, while fibrotic liver or a tumorin soft tissue, such as liver, is stiff. Noninvasive imaging techniquescan measure the elastic properties of soft tissue such as liver.Ultrasound and magnetic resonance are two modalities for measuringelasticity of soft tissue. Transient elastography, e.g., usingFIBROSCAN® Vibration Controlled Transient Elastrography (by echosens,France, is an ultrasound method and has demonstrated that liverstiffness progression is associated with clinical outcomes in PSC.Therefore, ‘progression to cirrhosis’ as assessed by FIBROSCAN® isanother alternative assessment for determining an effective amount.Magnetic Resonance Elastography (MRE) is a non-invasive imagingtechnique that may be used in subjects to assess liver stiffness toevaluate the impact of anti-α4β7 antibodies on inflammation andfibrosis. Other modalities include strain imaging, acoustic shear waveimaging and supersonic shear imaging.

In one embodiment, an assessment of the extent of liver fibrosis ismeasured using transient elastography. In another embodiment, anassessment of the extent of liver fibrosis is measured using magneticresonance elastography. An effective amount of anti-α4β7 antibody can bean amount sufficient to maintain, improve or normalize the elasticity ofthe liver. In some embodiments, an effective amount of anti-α4β7antibody reduces liver stiffness at least 10%, at least 20%, at least30%, at least 40%, at least 50% or at least 60% from the amount ofstiffness prior to treatment or since the prior dose of anti-α4β7antibody.

In other embodiments, an effective amount of an anti-α4β7 antibody canbe an amount sufficient to reduce morbidity or mortality of PSC. Forexample, an effective amount of an anti-α4β7 antibody can be an amountsufficient to provide one or more of the following effects: a reductionof or limit of the number of episodes of cholangitis, an increase in theperiod of time between episodes of cholangitis, a reduction in thenumber of strictures, a reduction of the likelihood of development ofportal hypertension or complications from hypertension, a reduction inhepatic osteodystrophy, such as a maintenance of bone density, and/or animprovement in pruritus. In some embodiments, an effective amount of ananti-α4β7 antibody can be an amount sufficient to reduce biliary diseaseburden, e.g. as measured on a magnetic resonance cholangiopancreatogram.

For therapy, an effective amount of anti-α4β7 antibody will besufficient to achieve the desired therapeutic (including prophylactic)effect (such as an amount sufficient to reduce or prevent α4β7integrin-mediated binding and/or signaling, thereby inhibiting leukocyteadhesion and infiltration and/or associated cellular responses in theliver and/or the bile ducts). An effective amount of an anti-α4β7antibody, e.g., an effective titer sufficient to maintain saturation,e.g., neutralization, of α4β7 integrin, can treat PSC.

An effective dose can be administered in a unit dose or multiple doses.The dosage can be determined by methods known in the art and can bedependent, for example, upon the individual's age, sensitivity,tolerance and overall well-being. A clinician or pharmacist of ordinaryskill can determine appropriate dosing using the guidance providedherein and conventional methods. For example, the levels of a marker inthe individual being treated can be measured (e.g., ALP) and dosing canbe adjusted to achieve the desired reduction or normalization of thelevel of the marker. Examples of modes of administration include topicalroutes such as nasal or inhalational or transdermal administration,enteral routes, such as through a feeding tube or suppository, andparenteral routes, such as intravenous, intramuscular, subcutaneous,intraarterial, intraperitoneal, or intravitreal administration. Suitabledosages for antibodies can be from about 0.1 mg/kg body weight to about10.0 mg/kg body weight per treatment, for example about 2 mg/kg to about7 mg/kg, about 3 mg/kg to about 6 mg/kg, or about 3.5 to about 5 mg/kg.In particular embodiments, the dose administered is about 0.3 mg/kg,about 0.5 mg/kg, about 1 mg/kg, about 2 mg/kg, about 3 mg/kg, about 4mg/kg, about 5 mg/kg, about 6 mg/kg, about 7 mg/kg, about 8 mg/kg, about9 mg/kg, or about 10 mg/kg. A dose of anti-α4β7 antibody can be in arange of 280 to 320 mg or antibody or consist of 300 mg of antibody. Adose of anti-α4β7 antibody can be in a range of 45 to 60 mg of antibodyor consist of 54 mg of antibody. A dose of anti-α4β7 antibody can be ina range of 95 to 115 mg of antibody or consist of 108 mg of antibody. Adose of anti-α4β7 antibody can be in a range of 150 to 175 mg ofantibody or consist of 160 or 165 mg of antibody. A dose of anti-α4β7antibody can consist of a double dose, e.g., 600 mg, 480 mg or 216 mg ofantibody.

In some embodiments, a formulation or composition comprising the anti-α4β7 antibody can comprise a mixture of a non-reducing sugar, e.g.,sucrose, an anti-α4β7 antibody and at least one free amino acid, and themolar ratio of non-reducing sugar to anti-α4β7 antibody (mole:mole) isgreater than 600:1 and the ratio of free amino acid to anti-α4β7antibody (mole:mole) is greater than 250:1. In liquid form, the liquidformulation or composition can comprise at least about 40 mg/ml to about80 mg/ml anti-α4β7 antibody, at least about 50-175 mM of one or moreamino acids, and at least about 6% to at least about 10% (w/v) sugar. Inanother embodiment, the formulation or composition is a lyophilizedformulation comprising a non-reducing sugar, an anti-α4β7 antibody,histidine, arginine and polysorbate 80, wherein the molar ratio ofnon-reducing sugar to anti-α4β7 antibody (mole:mole) in the formulationis greater than 600:1 and the molar ratio of arginine to anti-α4β7antibody (mole:mole) in the formulation is greater than 250:1. In liquidform, e.g., thawed, prior to lyophilization or after reconstitution froma dry state, a formulation can comprise 60 mg/mL anti-α4β7 antibody, 50mM histidine, 125 mM arginine, 10% sucrose, 0.06% polysorbate 80, pH6.3.

In other embodiments, a liquid formulation or composition comprising theanti- α4β7 antibody can comprise a mixture of anti-α4β7 antibody,citrate, histidine, arginine and polysorbate 80. In other embodiments, astable liquid pharmaceutical formulation for use in the treatment cancomprise anti-α4β7 antibody, citrate, histidine, arginine andpolysorbate 80. A liquid formulation or composition of anti-α4β7antibody can have an anti-α4β7 antibody concentration of greater than100 mg/ml, such as 120 to 180 mg/ml, 150 to 175 mg/ml or about 160 mg/ml(+/−5%). In an embodiment, the formulation or composition comprises atleast about 140 mg/ml or about 150 mg/ml to about 170 mg/ml, forexample, about 160 mg/ml of an anti-α4β7 antibody, a buffering agent(e.g., histidine), at least about 5 mM citrate to no more than 30 mMcitrate and a free amino acid (e.g., arginine). In another embodiment,the formulation or composition comprises at least about 160 mg/ml of ananti-α4β7 antibody, a buffering agent (e.g., histidine), at least about5 mM citrate, polysorbate 80 (e.g. 0.2% PS80), and a free amino acid(e.g., arginine). In an embodiment, a liquid formulation or compositioncan comprise 160 mg/mL anti-α4β7 antibody, 125 mM arginine, 50 mMhistidine, 25 mM citrate, and 0.2% PS80 (Protein Molar Ratio 1.5).Formulations of anti-α4β7 antibody are described in PCT publicationsWO2012151248 and WO2012151247, the entire contents of which areincorporated by this reference.

The final dosage form, e.g., after dilution of an anti-α4β7 antibody,e.g., reconstituted from a lyophilized formulation (e.g., then dilutedin a saline, Ringer's solution or 5% dextrose infusion system) of theanti-α4β7 antibody can be about 0.5 mg/ml to about 5 mg/ml foradministration. The final dosage form may be at a concentration ofbetween about 1.0 mg/ml to about 1.4 mg/ml, about 1.0 mg/ml to about 1.3mg/ml, about 1.0 mg/ml to about 1.2 mg/ml, about 1.0 to about 1.1 mg/ml,about 1.1 mg/ml to about 1.4 mg/ml, about 1.1 mg/ml to about 1.3 mg/ml,about 1.1 mg/ml to about 1.2 mg/ml, about 1.2 mg/ml to about 1.4 mg/ml,about 1.2 mg/ml to about 1.3 mg/ml, or about 1.3 mg/ml to about 1.4mg/ml. The final dosage form may be at a concentration of about 0.6mg/ml, 0.8 mg/ml, 1.0 mg/ml, 1.1 mg/ml, about 1.2 mg/ml, about 1.3mg/ml, about 1.4 mg/ml, about 1.5 mg/ml, about 1.6 mg/ml, about 1.8mg/ml or about 2.0 mg/ml. In one embodiment, the total dose is 180 mg.In another embodiment, the total dose is 300 mg. A 300 mg anti-α4β7antibody dose can be diluted into a larger volume, e.g., 250 ml, saline,Ringer's solution or 5% dextrose solution for administration, e.g.,intravenous administration.

In some aspects, the dosing regimen has two phases, an induction phaseand a maintenance phase. In the induction phase, the antibody orantigen-binding fragment thereof is administered in a way that quicklyprovides an effective amount of the antibody or antigen binding fragmentthereof suitable for certain purposes, such as inducing immune toleranceto the antibody or antigen-binding fragment thereof or for inducing aclinical response and ameliorating primary sclerosing cholangitissymptoms. A patient can be administered an induction phase treatmentwhen first being treated by an anti-α4β7 antibody, when being treatedafter a long absence from therapy, e.g., more than three months, morethan four months, more than six months, more than nine months, more thanone year, more than eighteen months or more than two years sinceanti-α4β7 antibody therapy or during maintenance phase of anti-α4β7antibody therapy if there has been a return of primary sclerosingcholangitis symptoms, e.g., a relapse from remission of disease, e.g.,an increase in ALP measurements (e.g., >ULN, e.g., >125 IU/L, >130IU/L, >135 IU/L, >140 IU/L or >145 IU/L) or an ALP measurement at least1.6 times ULN. In some embodiments, the induction phase regimen resultsin a higher mean trough serum concentration, e.g., the concentrationjust before the next dose, than the mean steady state trough serumconcentration maintained during the maintenance regimen.

In the maintenance phase, the antibody or antigen-binding fragmentthereof is administered in a way that continues the response achieved byinduction therapy with a stable level of antibody or antigen-bindingfragment thereof. A maintenance regimen can prevent return of symptomsor relapse of primary sclerosing cholangitis. A maintenance regimen canprovide convenience to the patient, e.g., be a simple dosing regimen orrequire infrequent trips for treatment. In some embodiments, themaintenance regimen can include administration of the anti-α4β7 antibodyor antigen-binding fragment thereof, e.g., in a formulation,composition, use and/or treatment method described herein, by a strategyselected from the group consisting of low dose, infrequentadministration, self-administration and a combination any of theforegoing.

In one embodiment, e.g., during an induction phase of therapy, thedosing regimen provides an effective amount of an anti-α4β7 antibody orantigen-binding fragment in a formulation, composition, use and/ortreatment method described herein for inducing remission of primarysclerosing cholangitis in a human patient. In some embodiments, theeffective amount of the anti-α4β7 antibody is sufficient to achieveabout 5 μg/ml to about 60 μg/ml, about 10 μg/ml to about 75 μg/ml, about15 μg/ml to about 45 μg/ml, about 20 μg/ml to about 30 μg/ml, or about25 μg/ml to about 35 μg/ml mean trough serum concentration of theanti-α4β7 antibody by the end of the induction phase. The duration ofinduction phase can be about three weeks, about four weeks, about fiveweeks, about six weeks, about seven weeks, about eight weeks, about tenweeks or about two to eight weeks of treatment, e.g., when the patientis beginning treatment with the anti-α4β7 antibody or has no anti-α4β7antibody in circulation (e.g., after a long elapsed time (such as 6months to two years) since the previous treatment. In some embodiments,the induction regimen can utilize a strategy selected from the groupconsisting of high dose, frequent administration, and a combination ofhigh dose and frequent administration of the anti-α4β7 antibody orantigen-binding fragment thereof, e.g., in a formulation or compositiondescribed herein. Induction dosing can be once, or a plurality of morethan one dose, e.g., at least two doses. During induction phase, a dosecan be administered once per day, every other day, twice per week, onceper week, once every ten days, once every two weeks or once every threeweeks. In some embodiments, the induction doses are administered withinthe first two weeks of therapy with the anti-α4β7 antibody. In oneembodiment, induction dosing can be once at initiation of treatment (day0) and once at about two weeks after initiation of treatment. In anotherembodiment, the induction phase duration is six weeks. In anotherembodiment, the induction phase duration is six weeks and a plurality ofinduction doses are administered during the first two weeks.

In some embodiments, e.g., when initiating treatment of a patient withsevere primary sclerosing cholangitis, the induction phase needs to havea longer duration than for patients with mild or moderate disease. Insome embodiments, the induction phase for a patient with a severedisease can have a duration of at least 6 weeks, at least 8 weeks, atleast 10 weeks, at least 12 weeks or at least 14 weeks. In oneembodiment, an induction dosing regimen for a patient with a severedisease can include a dose at week 0 (initiation of treatment), a doseat week 2 and a dose at week 6. In another embodiment, an inductiondosing regimen for a patient with a severe disease can comprise a doseevery week for four weeks or a dose at week 0 (initiation of treatment),a dose at week 2 and a dose at week 4. In another embodiment, aninduction dosing regimen for a patient with a severe disease cancomprise a dose at week 0 (initiation of treatment), a dose at week 2, adose at week 6 and a dose at week 10.

In one embodiment, e.g., during a maintenance phase of therapy, thedosing regimen maintains a mean steady state trough serum concentration,e.g., the plateau concentration just before the next dose, of about 5 toabout 25 μg/mL, about 7 to about 20 μg/mL, about 5 to about 10 μg/mL,about 10 to about 20 μg/mL, about 15 to about 25 μg/mL, about 15 toabout 40 μg/mL, about 20 to about 45 μg/mL or about 9 to about 13 μg/mLof anti-α4β7 antibody. In another embodiment, the dosing regimen e.g.,during a maintenance phase of therapy, maintains a mean steady statetrough serum concentration of about 10 to about 50 μg/mL, about 20 toabout 30 μg/mL, about 20 to about 55 μg/mL, about 30 to about 45 μg/mL,about 45 to about 55 μg/mL or about 35 to about 40 μg/mL of anti-α4β7antibody. In some embodiments, treatment with an effective amount ofanti-α4β7 antibody maintains a mean serum trough concentration ofanti-α4β7 antibody in a range of 20 to 100 μg/mL, 25 to 75 μg/mL or 30to 60 μg/mL. The amount of anti-α4β7 antibody can be measured using anagent which specifically detects the antibody. For example, ananti-idiotypic antibody raised from the anti-α4β7 antibody can be usedin assays, such as ELISA assays, to detect and measure the amount ofanti-α4β7 antibody in serum.

The dose can be administered twice per week, once per week, once every10 days, once every 2 weeks, once every 3 weeks, once every 4 weeks,once every month, once every 6 weeks, once every 8 weeks, once every 10weeks or once every 4 to 12 weeks. A higher or more frequent dose, e.g.,once or twice per week, once every 2 weeks, once every 3 weeks or onceevery 4 weeks, can be useful for inducing remission of active disease orfor treating a new patient, e.g., for inducing tolerance to the anti-α4β7 antibody. A less frequent dose, e.g., once every 4 weeks, onceevery 5 weeks, once every 6 weeks, once every 8 weeks, once every 10weeks or once every 4 to 12 weeks, can be useful for preventativetherapy, e.g., to maintain remission of a patient with chronic disease.In one aspect, the treatment regimen is treatment at day 0, about week2, about week 6 and every 4 or 8 weeks thereafter. In one embodiment,the maintenance regimen includes a dose of anti- α4β7 antibody every 4weeks. In one embodiment, the maintenance regimen includes a dose ofanti- α4β7 antibody every 8 weeks. In an embodiment, wherein a patienton a one dose every eight weeks maintenance regimen experiences a returnof one or more disease symptoms, e.g., has a relapse, the dosingfrequency can be increased, e.g., to once every 4 weeks.

The dose can be administered to the patient in about 20 minutes, about25 minutes, about 30 minutes, about 35 minutes, or about 40 minutes,e.g., as a continuous infusion, e.g., intravenously. The dose can beself-administered, e.g., by self-injection, such as a subcutaneousinjection.

The dosing regimen can be optimized to induce a clinical response andclinical remission in the primary sclerosing cholangitis of the patient.In some embodiments, the dosing regimen does not alter the ratio of CD4to CD8 in cerebrospinal fluid of patients receiving treatment.

In some aspects, a durable clinical remission, for example, a clinicalremission which is sustained through at least two, at least three, atleast four visits with a caretaking physician within a six month or oneyear period after beginning treatment may be achieved with an optimizeddosing regimen.

In some aspects, a durable clinical response, for example, a clinicalresponse which is sustained for at least 6 months, at least 9 months, atleast a year, at least 18 months after the start of treatment, may beachieved with an optimized dosing regimen.

In some embodiments, the dosing regimen for anti-α4β7 antibody therapycomprises an initial dose of anti-α4β7 antibody of 100 to 900 mg, 150 to500 mg or 250 to 350 mg, one to four subsequent doses of 100 to 900 mg,150 to 600 mg or 200 to 350 mg at about one to three week intervalswithin about one to ten weeks after the initial dose, and, thereafter(e.g., after the induction, initial higher doses), further subsequentdoses of 40 to 500 mg or 50 to 350 mg at about two to twelve weekintervals, e.g., in a maintenance phase regimen. In some embodiments,the anti-α4β7 antibody is vedolizumab. In some embodiments, all dosesare administered intravenously. In some embodiments, all doses areadministered subcutaneously. In some embodiments, the initial higherdoses (i.e., induction phase) are administered intravenously and thesubsequent lower, long interval doses (i.e., maintenance phase) areadministered subcutaneously. In some embodiments, the doses for eachadministration are provided in one article of manufacture, e.g, a singlevial or syringe. In other embodiments the doses are provided in morethan one article of manufacture, e.g, two or three vials or syringes.

In one embodiment, the dosing regimen comprises an initial dose of 300mg, a second subsequent dose of 300 mg about two weeks after the initialdose, a third subsequent dose of 300 mg at about six weeks after theinitial dose, followed by a fourth and subsequent doses of 300 mg everyfour weeks or every eight weeks after the third subsequent dose. In someembodiments, the anti-α4β7 antibody is vedolizumab. In some embodiments,all doses are administered intravenously.

In one embodiment, the dosing regimen for anti-α4β7 antibody therapycomprises an initial dose of anti-α4β7 antibody of 300 mg, a secondsubsequent dose of 300 mg about two weeks after the initial dose, athird subsequent dose of 108 mg at about six weeks after the initialdose, followed by a fourth and subsequent doses of 108 mg every week,every two weeks, every three weeks, or every four weeks after the thirdsubsequent dose. In some embodiments, the anti-α4β7 antibody isvedolizumab. In some embodiments, the induction doses are administeredintravenously and the maintenance doses are administered subcutaneously.

In one embodiment, the dosing regimen for anti-α4β7 antibody therapycomprises an initial dose of anti-α4β7 antibody of 160 mg, a secondsubsequent dose of 160 mg about one week after the initial dose, a thirdsubsequent dose of 160 mg at about two weeks after the initial dose, afourth subsequent dose of 108 mg at about four weeks after the initialdose, followed by a fifth and subsequent doses of 108 mg every week,every two weeks, every three weeks, or every four weeks after the thirdsubsequent dose. In some embodiments, the anti-α4β7 antibody isvedolizumab. In some embodiments, all doses are administeredsubcutaneously.

In some embodiments, the method of treatment, dose or dosing regimenreduces the likelihood that a patient will develop a HAHA response tothe anti-α4β7 antibody. The development of HAHA, e.g., as measured byantibodies reactive to the anti-α4β7 antibody, e.g., antibodies in apatient serum sample, can increase the clearance of the anti-α4β7antibody, e.g., reduce the serum concentration of the anti-α4β7antibody, e.g., lowering the number of anti-α4β7 antibody bound to α4β7integrin, thus making the treatment less effective. In some embodiments,to prevent HAHA, the patient can be treated with an induction regimenfollowed by a maintenance regimen. In some embodiments, there is nobreak between the induction regimen and the maintenance regimen. In someembodiments, the induction regimen comprises administering a pluralityof doses of anti-α4β7 antibody to the patient. To prevent HAHA, thepatient can be treated with a high initial dose, e.g., at least 1.5mg/kg, at least 2 mg/kg, at least 2.5 mg/kg, at least 3 mg/kg, at least5 mg/kg, at least 8 mg/kg, at least 10 mg/kg or about 2 to about 6mg/kg, or frequent initial administrations, e.g., about once per week,about once every two weeks or about once every three weeks, of thestandard dose when beginning therapy with an anti-α4β7 antibody. In someembodiments, the method of treatment maintains at least 30%, at least40%, at least 50%, at least 60%, at least 70%, at least 80%, at least90% or at least 95% of patients as HAHA-negative. In other embodiments,the method of treatment maintains patients as HAHA-negative for at least6 weeks, at least 10 weeks at least 15 weeks, at least six months, atleast 1 year, at least 2 years, or for the duration of therapy. In someembodiments, the patients, or at least 30%, at least 40%, at least 50%or at least 60% of patients who develop HAHA maintain a low titer, e.g.,<125, of anti-α4β7 antibody. In an embodiment, the method of treatmentmaintains at least 70% of patients as HAHA-negative for at least 12weeks after beginning therapy with an anti-α4β7 antibody. In anembodiment, the method of treatment maintains at least 80% of patientsas with no more than two consecutive samples, e.g., samples from twoconsecutive visits, testing HAHA-positive after beginning therapy withan anti-α4β7 antibody.

The formulation, composition or dose may be administered to anindividual (e.g., a human) alone or in conjunction with another agent. Aformulation, composition or dose of the invention can be administeredbefore, along with or subsequent to administration of the additionalagent. In one embodiment, more than one formulation or therapeutic agentwhich inhibits the binding of α4β7 integrin to its ligands isadministered. In such an embodiment, an agent, e.g., a monoclonalantibody, such as an anti-MAdCAM (e.g., anti-MAdCAM-1) or an anti-VCAM-1monoclonal antibody can be administered. In another embodiment, theadditional agent inhibits the binding of leukocytes to an endothelialligand in a pathway different from the α4β7 pathway. Such an agent caninhibit the binding, e.g. of chemokine (C-C motif) receptor 9(CCR9)-expressing lymphocytes to thymus expressed chemokine (TECK orCCL25) or an agent which prevents the binding of LFA-1 to intercellularadhesion molecule (ICAM). For example, an anti-TECK or anti-CCR9antibody or a small molecule CCR9 inhibitor, such as inhibitorsdisclosed in PCT publication WO03/099773 or WO04/046092, or anti-ICAM-1antibody or an oligonucleotide which prevents expression of ICAM, isadministered in addition to a formulation, dose or composition of thepresent invention. In yet another embodiment, an additional activeingredient (e.g., an anti-inflammatory compound, such as sulfasalazine,azathioprine, 6-mercaptopurine, 5-aminosalicylic acid containinganti-inflammatories, another non-steroidal anti-inflammatory compound, asteroidal anti-inflammatory compound, or antibiotics commonlyadministered for control of IBD (e.g. ciprofloxacin, metronidazole), oranother biologic agent (e.g. TNF alpha antagonists) can be administeredin conjunction with a formulation, composition or dose of the presentinvention. In an embodiment, a bile analog, e.g., ursodeoxycholic acid,can be administered in conjunction with a formulation, composition ordose of the anti-α4β7 antibody.

In an embodiment, the dose of the co-administered medication can bedecreased over time during the period of treatment by the formulation,composition or dose comprising the anti-α4β7 antibody. For example, apatient being treated with a steroid (e.g. prednisone, prednisolone) atthe beginning, or prior to, treating with the anti-α4β7 antibodyformulation, composition or dose would undergo a regimen of decreasingdoses of steroid beginning as early as 6 weeks of treatment with theanti-α4β7 antibody formulation. The steroid dose will be reduced byabout 25% within 4-8 weeks of initiating tapering, by 50% at about 8-12weeks and 75% at about 12-16 weeks of tapering during treatment with theanti-α4β7 antibody formulation, composition or dose. In one aspect, byabout 16-24 weeks of treatment with the anti-α4β7 antibody formulation,composition or dose, the steroid dose can be eliminated. In anotherexample, a patient being treated with an anti-inflammatory compound,such as 6-mercaptopurine at the beginning, or prior to, treating withthe anti-α4β7 antibody formulation, composition or dose would undergo aregimen of decreasing doses of anti-inflammatory compound similar to thetapering regimen for steroid dosing as noted above.

In one embodiment, the method comprises administering an effectiveamount of a pharmaceutical composition of an anti-α4β7 antibody to apatient, e.g., a patient suffering from PSC. The pharmaceuticalcomposition may be administered when the patient is in a fed or fastedcondition. If the pharmaceutical composition is in a solid, e.g., drystate, the process of administration can comprise a step of convertingthe pharmaceutical composition to a liquid state. In one aspect, a drypharmaceutical composition can be reconstituted, e.g., by a liquid, suchas water, isotonic saline or Ringer's solution , for use in injection,e.g. intravenous, intramuscular or subcutaneous injection. In anembodiment, if the pharmaceutical composition is supplied as a liquid,administration can proceed directly, e.g, for use in injection, e.g.intravenous, intramuscular or subcutaneous injection. In another aspect,a solid or dry pharmaceutical composition can be administered topically,e.g., in a patch, cream, aerosol or suppository.

In another aspect, the pharmaceutical composition of an anti-α4β7antibody is a stable liquid formulation or composition, e.g., as asolution comprising anti-α4β7 antibody in a vial, cartridge, prefilledsyringe or an autoinjector. The stable liquid formulation can comprise ahigh concentration of anti-α4β7 antibody, such as at least 60 mg/ml, atleast 90 mg/ml, at least 120 mg/ml, at least 150 mg/ml, at least155mg/ml, at least 160 mg/ml, at least 165 mg/ml, 130 mg/ml to 180mg/ml, 50 to 180 mg/ml, 60 to 70 mg/ml, 100 to 200 mg/ml, 145 to 175mg/ml, or about 160 mg/ml. A stable liquid formulation or compositioncomprising anti-α4β7 antibody can be administered subcutaneously,intravenously, or intramuscularly. A stable liquid formulationcomprising anti-α4β7 antibody can be self-administered.

In some embodiments, treatment with an anti-α4β7 antibody does not alterthe ratio of CD4:CD8 lymphocytes. CD4:CD8 ratios can be measured inblood, lymph node aspirate, and cerebro-spinal fluid (CSF). The CSFCD4+:CD8+lymphocyte ratios in healthy individuals are typically greaterthan or equal to about 1. (Svenningsson et al., J. Neuroimmunol.1995;63:39-46; Svenningsson et al., Ann Neurol. 1993; 34:155-161). Animmunomodulator can alter the CD4:CD8 ratio to less than 1.

In some embodiments, treatment with an anti-α4β7 antibody lengthens theamount of time to a PSC-related outcome or complication selected fromthe group consisting of death, liver failure, liver transplantation,ascites, hepatic encephalopathy, development of varices, jaundice,variceal bleeding, cholangiocarcinoma, hepatocellular carcinoma,evidence of cirrhosis, and colorectal cancer. In an embodiment, thetreatment decreases the chance of progression to cirrhosis.

In one aspect, the invention relates to a method of treating PSC in asubject with an effective amount of an anti-α4β7 antibody, wherein saidtreatment does not cause an adverse event such as, for example,hepatotoxicity, progressive multifocal leukoencephalopathy (PML),cholangiocarcinoma, complications of portal hypertension (such asascites, hepatic encephalopathy, development of varices, or jaundice),infusion-related reactions, infection, leucopenia, lymphopenia,colorectal cancer, acute respiratory failure, acute respiratory distresssyndrome, Torsade de pointes, ventricular fibrillation, ventriculartachycardia, malignant hypertension, convulsive seizure,agranulocytosis, aplastic anemia, toxic epidermal necrolysis,Stevens-Johnson syndrome, hepatic necrosis, acute liver failure,anaphylactic shock, acute renal failure, pulmonary hypertension,pulmonary fibrosis, confirmed or suspected endotoxin shock, confirmed orsuspected transmission of infectious agent by a medicinal product,neuroleptic malignant syndrome, malignant hyperthermia, spontaneousabortion, stillbirth, or fetal death. In some embodiments, a method oftreating PSC in a subject with an effective amount of an anti-α4β7antibody does not cause an adverse event such as ALT or AST elevated,e.g., in 2 consecutive measurements, >5 times their ULN or >5 timestheir baseline levels prior to treatment. In some embodiments, a methodof treating PSC in a subject with an effective amount of an anti-α4β7antibody does not cause an adverse event such as ALT or AST elevated >3times their ULN or >3 times their baseline levels prior to treatment andthe total bilirubin is >2 times its ULN or >2 times its baseline priorto treatment.

α4β7 Antibodies

Anti-α4β7 antibodies (anti-α4β7 integrin antibodies) suitable for use inthe formulations, compositions, uses, methods and/or doses includeantibodies from any desired source, such as fully human antibodies,murine antibodies, rabbit antibodies and the like, and any desiredengineered antibodies, such as chimeric antibodies, humanizedantibodies, and the like. Antigen-binding fragments of any of thesetypes of antibodies, such as Fab, Fv, scFv, Fab′ and F(ab′)₂ fragments,are also suitable for use in the formulations, compositions, uses,methods and/or doses.

The anti-α4β7 antibody can bind to an epitope on the α4 chain (e.g.,humanized MAb 21.6 (Bendig et al., U.S. Pat. No. 5,840,299), on the β7chain (e.g., FIB504 or a humanized derivative (e.g., Fong et al., U.S.Pat. No. 7,528,236)), or to a combinatorial epitope formed by theassociation of the α4 chain with the (37 chain. In one aspect, theantibody binds a combinatorial epitope on the α4β7 complex, but does notbind an epitope on the α4 chain or the β7 chain unless the chains are inassociation with each other. The association of α4 integrin with β7integrin can create a combinatorial epitope for example, by bringinginto proximity residues present on both chains which together comprisethe epitope or by conformationally exposing on one chain, e.g., the α4integrin chain or the β7 integrin chain, an epitopic binding site thatis inaccessible to antibody binding in the absence of the properintegrin partner or in the absence of integrin activation. In anotheraspect, the anti-α4β7 antibody binds both the α4 integrin chain and theβ7 integrin chain, and thus, is specific for the α4β7 integrin complex.Such antibodies can bind α4β7 but not bind α4β1, and/or not bindα_(E)β7, for example. In another aspect, the anti-α4β7 antibody binds tothe same or substantially the same epitope as the Act-1 antibody(Lazarovits, A. I. et al., J. Immunol., 133(4): 1857-1862 (1984),Schweighoffer et al., J. Immunol., 151(2): 717-729, 1993; Bednarczyk etal., J. Biol. Chem., 269(11): 8348-8354, 1994). Murine ACT-1 Hybridomacell line, which produces the murine Act-1 monoclonal antibody, wasdeposited under the provisions of the Budapest Treaty on Aug. 22, 2001,on behalf Millennium Pharmaceuticals, Inc., 40 Landsdowne Street,Cambridge, Mass. 02139, U.S.A., at the American Type Culture Collection,10801 University Boulevard, Manassas, Va. 20110-2209, U.S.A., underAccession No. PTA-3663. In another aspect, the anti-α4β7 antibody is ahuman antibody or an α4β7 binding protein using the CDRs provided inU.S. Patent Application Publication No. 2010/0254975.

In one aspect, the anti-α4β7 antibody inhibits binding of α4β7 to one ormore of its ligands (e.g. the mucosal addressin, e.g., MAdCAM (e.g.,MAdCAM-1), fibronectin, and/or vascular addressin (VCAM)). PrimateMAdCAMs are described in the PCT publication WO 96/24673, the entireteachings of which are incorporated herein by this reference. In anotheraspect, the anti-α4β7 antibody inhibits binding of α4β7 to MAdCAM (e.g.,MAdCAM-1) and/or fibronectin without inhibiting the binding of VCAM.

In one aspect, the anti-α4β7 antibodies for use in the formulations,compositions, uses, doses and/or treatment methods described herein arehumanized versions of the mouse Act-1 antibody. Suitable methods forpreparing humanized antibodies are well-known in the art. Generally, thehumanized anti-α4β7 antibody will contain a heavy chain that containsthe 3 heavy chain complementarity determining regions (CDRs, CDR1, SEQID NO:8, CDR2, SEQ ID NO:9 and CDR3, SEQ ID NO:10) of the mouse Act-1antibody and suitable human heavy chain framework regions; and alsocontain a light chain that contains the 3 light chain CDRs (CDR1, SEQ IDNO:11, CDR2, SEQ ID NO:12 and CDR3, SEQ ID NO:13) of the mouse Act-1antibody and suitable human light chain framework regions. The humanizedAct-1 antibody can contain any suitable human framework regions,including consensus framework regions, with or without amino acidsubstitutions. For example, one or more of the frame work amino acidscan be replaced with another amino acid, such as the amino acid at thecorresponding position in the mouse Act-1 antibody. The human constantregion or portion thereof, if present, can be derived from the κ or λ,light chains, and/or the γ (e.g., γ, γ2, γ3, γ4), μ, α (e.g., α1, α2), δor ε heavy chains of human antibodies, including allelic variants. Aparticular constant region (e.g., IgG1), variant or portions thereof canbe selected in order to tailor effector function. For example, a mutatedconstant region (variant) can be incorporated into a fusion protein tominimize binding to Fc receptors and/or ability to fix complement (seee.g., Winter et al., GB 2,209,757 B; Morrison et al., WO 89/07142;Morgan et al., WO 94/29351, Dec. 22, 1994). Humanized versions of Act-1antibody were described in PCT publications nos. WO98/06248 andWO07/61679, the entire teachings of each of which are incorporatedherein by this reference.

In another aspect, the humanized anti-α4β7 antibody for use in theformulations, compositions, uses, doses and/or treatment methodscomprises a heavy chain variable region comprising amino acids 20 to 140of SEQ ID NO:2, and a light chain variable region comprising amino acids20 to 131 of SEQ ID NO:4 or amino acids 21 to 132 of SEQ ID NO:5. Ifdesired, a suitable human constant region(s) can be present. Forexample, the humanized anti-α4β7 antibody can comprise a heavy chainthat comprises amino acids 20 to 470 of SEQ ID NO:2 and a light chaincomprising amino acids 21 to 239 of SEQ ID NO:5. In another example, thehumanized anti-α4β7 antibody can comprise a heavy chain that comprisesamino acids 20 to 470 of SEQ ID NO:2 and a light chain comprising aminoacids 20 to 238 of SEQ ID NO:4. The humanized anti-α4β7 antibody can beencoded by a heavy chain nucleic acid sequence of SEQ ID NO:1 and alight chain nucleic sequence of SEQ ID NO:3. The humanized light chainof vedolizumab (e.g., Chemical Abstract Service (CAS, American ChemicalSociety) Registry number 943609-66-3), with two mouse residues switchedfor human residues, is more human than the light chain of LDP-02. Inaddition, LDP-02 has the somewhat hydrophobic, flexible alanine 114 anda hydrophilic site (Aspartate 115) that is replaced in vedolizumab withthe slightly hydrophilic hydroxyl-containing threonine 114 andhydrophobic, potentially inward facing valine 115 residue.

Further substitutions to the humanized anti-α4β7 antibody sequence canbe, for example, mutations to the heavy and light chain frameworkregions, such as a mutation of isoleucine to valine on residue 2 of SEQID NO:14; a mutation of methionine to valine on residue 4 of SEQ IDNO:14; a mutation of alanine to glycine on residue 24 of SEQ ID NO:15; amutation of arginine to lysine at residue 38 of SEQ ID NO:15; a mutationof alanine to arginine at residue 40 of SEQ ID NO:15; a mutation ofmethionine to isoleucine on residue 48 of SEQ ID NO:15; a mutation ofisoleucine to leucine on residue 69 of SEQ ID NO:15; a mutation ofarginine to valine on residue 71 of SEQ ID NO:15; a mutation ofthreonine to isoleucine on residue 73 of SEQ ID NO:15; or anycombination thereof; and replacement of the heavy chain CDRs with theCDRs (CDR1, SEQ ID NO:8, CDR2, SEQ ID NO:9 and CDR3, SEQ ID NO:10) ofthe mouse Act-1 antibody; and replacement of the light chain CDRs withthe light chain CDRs (CDR1, SEQ ID NO:11, CDR2, SEQ ID NO:12 and CDR3,SEQ ID NO:13) of the mouse Act-1 antibody.

In some embodiments, the anti-α4β7 antibody, e.g., a humanized anti-α4β7antibody for use in the formulation, compositions, uses, doses and/ortreatment methods comprises a heavy chain variable region that has about95%, 96%, 97%, 98%, or 99% sequence identity to amino acids 20 to 140 ofSEQ ID NO:2, and a light chain variable region that has about 95%, 96%,97%, 98%, or 99% sequence identity to amino acids 20 to 131 of SEQ IDNO:4 or amino acids 21 to 132 of SEQ ID NO:5. Amino acid sequenceidentity can be determined using a suitable sequence alignmentalgorithm, such as the Lasergene system (DNASTAR, Inc., Madison, Wis.),using the default parameters. In an embodiment, the anti-α4β7 antibodyfor use in the formulation, compositions, uses, methods and/or doses isvedolizumab (CAS, American Chemical Society, Registry number943609-66-3).

Other α4β7 antibodies may also be used in the formulations,compositions, uses, methods and/or dosing regimes described herein. Forexample, the α4β7 antibodies described in US 2010/0254975 (Amgen, Inc.)or the β7 antibodies described in WO2006026759 (Genentech, Inc.),incorporated by reference herein in its entirety, are suitable for usein the formulations, compositions, uses, doses and/or methods oftreating primary sclerosing cholangitis in an individual.

The anti-α4β7 antibody can be produced by expression of nucleic acidsequences encoding each chain in living cells, e.g., cells in culture. Avariety of host-expression vector systems may be utilized to express theantibody molecules of the invention. Such host-expression systemsrepresent vehicles by which the coding sequences of interest may beproduced and subsequently purified, but also represent cells which may,when transformed or transfected with the appropriate nucleotide codingsequences, express an anti-α4β7 antibody in situ. These include but arenot limited to microorganisms such as bacteria (e.g., E. coli, B.subtilis) transformed with recombinant bacteriophage DNA, plasmid DNA orcosmid DNA expression vectors containing antibody coding sequences;yeast (e.g., Saccharomyces, Pichia) transformed with recombinant yeastexpression vectors containing antibody coding sequences; insect cellsystems infected with recombinant virus expression vectors (e.g.,baculovirus) containing antibody coding sequences; plant cell systemsinfected with recombinant virus expression vectors (e.g., cauliflowermosaic virus, CaMV; tobacco mosaic virus, TMV) or transformed withrecombinant plasmid expression vectors (e.g., Ti plasmid) containingantibody coding sequences; or mammalian cell systems (e.g., COS, CHO,BHK, 293, 3T3, NSO cells) harboring recombinant expression constructscontaining promoters derived from the genome of mammalian cells (e.g.,metallothionein promoter) or from mammalian viruses (e.g., theadenovirus late promoter; the vaccinia virus 7.5K promoter). Forexample, mammalian cells such as Chinese hamster ovary cells (CHO), inconjunction with a vector such as the major intermediate early genepromoter element from human cytomegalovirus is an effective expressionsystem for antibodies (Foecking et al., Gene 45:101 (1986); Cockett etal., Bio/Technology 8:2 (1990)).

In bacterial systems, a number of expression vectors may beadvantageously selected depending upon the use intended for the antibodymolecule being expressed. For example, when a large quantity of such aprotein is to be produced, for the generation of pharmaceuticalcompositions of an antibody molecule, vectors which direct theexpression of high levels of fusion protein products that are readilypurified may be desirable. Such vectors include, but are not limited, tothe E. coli expression vector pUR278 (Ruther et al., EMBO J. 2:1791(1983)), in which the antibody coding sequence may be ligatedindividually into the vector in frame with the lac Z coding region sothat a fusion protein is produced; pIN vectors (Inouye & Inouye, NucleicAcids Res. 13:3101-3109 (1985); Van Heeke & Schuster, J. Biol. Chem.24:5503-5509 (1989)); and the like. pGEX vectors may also be used toexpress foreign polypeptides as fusion proteins with glutathioneS-transferase (GST). In general, such fusion proteins are soluble andcan easily be purified from lysed cells by adsorption and binding tomatrix glutathione-agarose beads followed by elution in the presence offree glutathione. The pGEX vectors are designed to include thrombin orfactor Xa protease cleavage sites so that the cloned target gene productcan be released from the GST moiety.

In an insect system, Autographa californica nuclear polyhedrosis virus(AcNPV) is used as a vector to express foreign genes. The virus grows inSpodoptera frugiperda cells. The antibody coding sequence may be clonedindividually into non-essential regions (for example the polyhedringene) of the virus and placed under control of an AcNPV promoter (forexample the polyhedrin promoter).

In mammalian host cells, a number of viral-based expression systems maybe utilized. In cases where an adenovirus is used as an expressionvector, the antibody coding sequence of interest may be ligated to anadenovirus transcription/translation control complex, e.g., the latepromoter and tripartite leader sequence. This chimeric gene may then beinserted in the adenovirus genome by in vitro or in vivo recombination.Insertion in a non-essential region of the viral genome (e.g., region E1or E3) will result in a recombinant virus that is viable and capable ofexpressing the antibody molecule in infected hosts (e.g., see Logan &Shenk, Proc. Natl. Acad. Sci. USA 81:355-359 (1984)). Specificinitiation signals may also be required for efficient translation ofinserted antibody coding sequences. These signals include the ATGinitiation codon and adjacent sequences. Furthermore, the initiationcodon must be in phase with the reading frame of the desired codingsequence to ensure translation of the entire insert. These exogenoustranslational control signals and initiation codons can be of a varietyof origins, both natural and synthetic. The efficiency of expression maybe enhanced by the inclusion of appropriate transcription enhancerelements, transcription terminators, etc. (see Bittner et al., Methodsin Enzymol. 153:51-544 (1987)).

In addition, a host cell strain may be chosen which modulates theexpression of the inserted sequences, or modifies and processes the geneproduct in the specific fashion desired. Such modifications (e.g.,glycosylation) and processing (e.g., cleavage) of protein products maybe important for the function of the protein. Different host cells havecharacteristic and specific mechanisms for the post-translationalprocessing and modification of proteins and gene products. Appropriatecell lines or host systems can be chosen to ensure the correctmodification and processing of the foreign protein expressed. To thisend, eukaryotic host cells which possess the cellular machinery forproper processing of the primary transcript, glycosylation, andphosphorylation of the gene product may be used. Such mammalian hostcells include but are not limited to Chinese hamster ovary (CHO), NSO,HeLa, VERY, baby hamster kidney (BHK), monkey kidney (COS), MDCK, 293,3T3, WI38, human hepatocellular carcinoma cells (e.g., Hep G2), breastcancer cell lines such as, for example, BT483, Hs578T, HTB2, BT20 andT47D, and normal mammary gland cell line such as, for example, CRL7030and Hs578Bst.

The glycosylation machinery of different cell types can produceantibodies with different glycosylation composition than in another celltype, or no glycosylation, as with bacterial cells. In one aspect, celltypes for production of the anti-α4β7 antibody are mammalian cells, suchas NS0 or CHO cells. In one aspect, the mammalian cells can comprise thedeletion of an enzyme involved in cell metabolism and the exogenous geneof interest can be operably linked to a replacement enzyme, e.g., in aconstruct or vector for introduction into the cells, e.g., bytransformation or transfection. The construct or vector with theexogenous gene confers to the cells which host the construct or vector aselection advantage to encourage production of the polypeptide encodedby the exogenous gene. In one embodiment, CHO cells are DG44 cells(Chasin and Urlaub (1980) PNAS USA 77:4216), comprising the deletion orinactivation of the dihydrofolate reductase gene. In another embodiment,CHO cells are CHO K1 cells comprising the deletion or inactivation ofthe glutamine synthase gene (see, e.g., U.S. Pat. Nos. 5,122,464 or5,827,739).

In one aspect, the anti-α4β7 antibody is vedolizumab. Vedolizumab IV(also called MLN0002, ENTYVIO™ or KYNTELES™) is a humanizedimmunoglobulin (Ig) G1 mAb directed against the human lymphocyteintegrin α4β7. The α4β7 integrin mediates lymphocyte trafficking to GImucosa and gut-associated lymphoid tissue (GALT) through adhesiveinteraction with mucosal addressin cell adhesion molecule-1 (MAdCAM-1),which is expressed on the endothelium of mesenteric lymph nodes and GImucosa. Vedolizumab binds the α4β7 integrin, antagonizes its adherenceto MAdCAM-1 and as such, impairs the migration of gut homing leukocytesinto GI mucosa. Without being bound by theory, it is believed thatvedolizumab may be used as a therapeutic agent for PSC by blockingT-cell recruitment and the ongoing inflammatory process by binding toα4β7 integrin expressed on circulating lymphocytes.

Pharmaceutical Compositions

The anti-α4β7 antibody can be administered to the individual as part ofa pharmaceutical or physiological composition for the treatment ofprimary sclerosing cholangitis (PSC). Such a composition can comprise anantibody or antigen-binding fragment thereof having binding specificityfor α4β7 integrin as described herein, and a pharmaceutically orphysiologically acceptable carrier. Pharmaceutical or physiologicalcompositions for co-therapy can comprise an antibody or antigen-bindingfragment having binding specificity for α4β7 integrin and one or moreadditional therapeutic agents. An antibody or antigen-binding fragmenthaving binding specificity for α4β7 integrin function and an additionaltherapeutic agent can be components of separate compositions which canbe mixed together prior to administration or administered separately.Formulation or composition will vary according to the route ofadministration selected (e.g., solution, emulsion, capsule). Suitablecarriers can contain inert ingredients which do not interact with theantibody or antigen-binding fragment and/or additional therapeuticagent. Standard pharmaceutical formulation techniques can be employed,such as those described in Remington's Pharmaceutical Sciences, MackPublishing Company, Easton, Pa. Suitable carriers for parenteraladministration include, for example, sterile water, physiologicalsaline, bacteriostatic saline (saline containing about 0.9% mg/ml benzylalcohol), phosphate-buffered saline, Hank's solution, Ringer's-lactateand the like. Methods for encapsulating compositions (such as in acoating of hard gelatin or cyclodextran) are known in the art (Baker, etal., “Controlled Release of Biological Active Agents”, John Wiley andSons, 1986). For inhalation, the agent can be solubilized and loadedinto a suitable dispenser for administration (e.g., an atomizer,nebulizer or pressurized aerosol dispenser).

For example, the pharmaceutical composition may comprise a mixture of anon-reducing sugar, an anti-α4β7 antibody and at least one free aminoacid, and the molar ratio of non-reducing sugar to anti-α4β7 antibody(mole:mole) is greater than 600:1. The pharmaceutical composition may bea liquid formulation or a dry formulation (e.g., lyophilized). Thepharmaceutical composition can also contain a buffering agent. In someembodiments, the non-reducing sugar is mannitol, sorbitol, sucrose,trehalose, or any combination thereof. The free amino acid of thepharmaceutical composition may be histidine, alanine, arginine, glycine,glutamic acid, or any combination thereof. The pharmaceuticalcomposition can comprise between about 50 mM to about 175 mM of freeamino acid. The pharmaceutical composition can comprise between about100 mM and about 175 mM of free amino acid. The ratio of free amino acidto antibody molar ratio can be at least 250:1. The pharmaceuticalcomposition can also contain a surfactant. The surfactant can bepolysorbate 20, polysorbate 80, a poloxamer, or any combination thereof.

The present invention will now be illustrated by the following Examples,which are not intended to be limiting in any way.

EXEMPLIFICATION Example 1 A Randomized, Global, Double-Blind,Placebo-Controlled, Parallel-Group Study to Evaluate the Efficacy andSafety of Vedolizumab IV for the Treatment of Primary SclerosingCholangitis

A phase 3 randomized, global, double-blind, parallel group, multicenterstudy will be done to assess the safety and efficacy of vedolizumab 300mg IV infusion over a 106 week treatment period in non-end stage PSCsubjects who also have inflammatory bowel disease (IBD). The study willinclude a 4-week Screening Period, a 106 week Treatment Period, and an18-week Follow-up Period starting at Week 102 or the last dose of studydrug for early withdrawal subjects. In addition, subjects willparticipate in a final safety visit at Week 120 and a follow-up safetysurvey at 6, 12, 18 and 24 months after the dose received at Week 102.Subjects who withdraw early from the study will be asked to undergo theend-of-treatment (EOT) assessments, including a liver biopsy (a liverbiopsy will only be requested if they have completed ≧12 monthstreatment)and will begin the 24-month follow-up safety survey startingfrom their last dose date).

Approximately 228 subjects with non-endstage PSC and underlying IBD willbe enrolled into the study. The subjects will be randomized into one of2 treatment groups in a 2:1 ratio, stratified by concomitant use ofursodeoxycholic acid (UDCA) at baseline and participation in themagnetic resonance elastography (MRE) substudy. The total percentage ofsubjects on UDCA will be limited to approximately 25% and these subjectswill be required to be at stable dose of up to 20 mg/kg UDCA for 8 weeksprior to the screening visit. If a subject needs to stop or change theirUDCA dose, prior to study entry, this must occur 8 weeks prior to theScreening.

The treatment groups will be:

A: vedolizumab IV 300 mg at Day 1 (week 0) and Week 2; followed byvedolizumab IV 300 mg every 4 weeks (Q4W) starting from Week 6 (Weeks 6,10, 14, 18, 22, 26, 30, 34, 38, 42, 46, 50, 54, 58, 62, 66, 70, 74, 78,82, 86, 90, 94, 98, and 102).

B: placebo IV at Day 1 (week 0) and Week 2; followed by placebo Q4Wstarting from Week 6 (Weeks 6, 10, 14, 18, 22, 26, 30, 34, 38, 42, 46,50, 54, 58, 62, 66, 70, 74, 78, 82, 86, 90, 94, 98, and 102).

An MRE substudy will be conducted at selected qualified sites, in amaximum of 114 subjects. This substudy will evaluate the effect ofvedolizumab on fibrosis by assessing liver stiffness by MRE. Onlysubjects from the pre-selected sites will be included in this study;these subjects will undergo an MRE assessment at Screening, Week 54 andWeek 106 or early termination (ET) visit, unless they havecontraindications to the procedure. All MRE results will be evaluated byan independent central reviewer.

An interim analysis will be conducted when 50% of the maximum number ofsubjects (114 across both groups) have completed the 106 week treatmentperiod or withdrawn from the study.

Pharmacogenomic analysis may be conducted to investigate thecontribution of genetic variance on drug response, for example, itsefficacy and safety.

The proposed study will also evaluate the effect of vedolizumab oninflammation, symptoms, biochemical markers, and other disease relatedclinical endpoints in non-endstage PSC subjects with underlying IBD.Pharmacokinetics (PK), pharmacodynamics (PD), and immunogenicity ofvedolizumab in subjects with non-endstage PSC will be evaluated.

The impact of vedolizumab on patient reported outcomes includesquantifying health-related quality of life (HRQOL), using the ChronicLiver Disease Questionnaire (CLDQ) and EuroQOL (EQ-5D-3L), and workproductivity, using the work productivity and activity impairmentquestionnaire (WPAI) in non-endstage PSC subjects.

For statistical analysis, the Full Analysis Set (FAS) will include allrandomized subjects who receive at least 1 dose of study drug. Subjectsin this set will be analyzed according to the treatment they wererandomized to receive.

The Safety Analysis Set will include all subjects who receive at least 1dose of study drug. Subjects in this set will be analyzed according tothe treatment they actually received.

The PK and PD evaluable population is defined as all subjects whoreceive at least 1 dose of study drug and have sufficient blood samplingto allow for PK and PD evaluation.

The time to event endpoints will be analyzed using a Cox proportionalhazard model with treatment group as a factor and stratified byrandomization stratum.

Example 2 Identification of Patients Suffering from PSC

Background: A rare cholestatic liver disease and often concomitant withinflammatory bowel disease (IBD), primary sclerosing cholangitis (PSC)is broadly coded in non-Read code-based databases, e.g., UK ClinicalPractice Research Datalink (CPRD GOLD). Within CPRD, Read codes for PSCand other forms of cholangitis are available. It is unknown if theseconditions are appropriately recorded in the CPRD.

Objectives: To assess information in patients with PRC diagnosis in CPRD

Methods: Patients with a Read code for PSC in 1988-2013 but without asecondary sclerosing cholangitis diagnosis any time were eligible. Weanalyzed patient characteristics before PSC diagnosis among those withat least one year data before and after the first diagnosis.

Results: 371 patients (mean age 54±18, men 58.2%) were identified withPSC, of whom 9.7%, 3.2%, and 0.5% also had Read codes for cholangitis,sclerosing cholangitis, and other cholangitis diagnosis, respectively.222 (59.8%) patients had at least one liver function test recorded. Thenumber of patients tested and the percentage of patients with abnormalresults were as follows: alkaline phosphatase (217, 87.1%), alaninetransaminase (178, 55.6%), γ-Glutamyl transpeptidase (156, 87.2%),aspartate transaminase (69, 42.0%), and total bilirubin (208, 15.9%).IBD (43.7%, mostly ulcerative colitis 37.2%) was the most common medicalhistory, followed by benign neoplasms (12.9%), cancers (5.9%, solid5.1%), biliary cirrhosis (2.2%), and liver transplantation (1.4%).

Conclusions: A variety of indicators reflected a PSC diagnosis in CPRDGOLD. Research in assessing potential miscoding of PSC and developingPSC coding algorithms is under way.

The previous description is provided to enable any person skilled in theart to practice the various aspects described herein. The previousdescription provides various examples of the subject technology, and thesubject technology is not limited to these examples. Variousmodifications to these aspects will be readily apparent to those skilledin the art, and the generic principles defined herein may be applied toother aspects. Thus, the claims are not intended to be limited to theaspects shown herein, but is to be accorded the full scope consistentwith the language claims, wherein reference to an element in thesingular is not intended to mean “one and only one” unless specificallyso stated, but rather “one or more.” Unless specifically statedotherwise, the term “some” refers to one or more. Headings andsubheadings, if any, are used for convenience only and do not limit theinvention.

SEQUENCE LISTING SEQ ID NO: 1gaattctcga gatcgatctc accatgggat ggagctgtat catcctatc ttggtagcaa    60cagctacagg tgtccactcc caggtgcaat tggtgcagtc tggggctgag gttaagaagc  120ctggggcttc agtgaaggtg tcctgcaagg gttctggcta caccttcacc agctactgga  180tgcattgggt gaggcaggcg cctggccaac gtctagagtg gatcggagag attgatcctt  240ctgagagtaa tactaactac aatcaaaaat tcaagggacg cgtcacattg actgtagaca  300tttccgctag cacagcctac atggagctct ccagcctgag atctgaggac actgcggtct  360actattgtgc aagagggggt tacgacggat gggactatgc tattgactac tggggtcaag  420gcaccctggt caccgtcagc tcagcctcca ccaagggccc atcggtcttc cccctggcac  480cctcctccaa gagcacctct gggggcacag cggccctggg ctgcctggtc aaggactact  540tccccgaacc ggtgacggtg tcgtggaact caggcgccct gaccagcggc gtgcacacct  600tcccggctgt cctacagtcc tcaggactct actccctcag cagcgtggtg accgtgccct  660ccagcagctt gggcacccag acctacatct gcaacgtgaa tcacaagccc agcaacacca  720aggtggacaa gaaagttgag cccaaatctt gtgacaaaac tcacacatgc ccaccgtgcc  780cagcacctga actcgcgggg gcaccgtcag tcttcctctt ccccccaaaa cccaaggaca  840ccctcatgat ctcccggacc cctgaggtca catgcgtggt ggtggacgtg agccacgaag  900accctgaggt caagttcaac tggtacgtgg acggcgtgga ggtgcataat gccaagacaa  960agccgcggga ggagcagtac aacagcacgt accgtgtggt cagcgtcctc accgtcctgc 1020accaggactg gctgaatggc aaggagtaca agtgcaaggt ctccaacaaa gccctcccag 1080cccccatcga gaaaaccatc tccaaagcca aagggcagcc ccgagaacca caggtgtaca 1140ccctgccccc atcccgggat gagctgacca agaaccaggt cagcctgacc tgcctggtca 1200aaggcttcta tcccagcgac atcgccgtgg agtgggagag caatgggcag ccggagaaca 1260actacaagac cacgcctccc gtgctggact ccgacggctc cttcttcctc tacagcaagc 1320tcaccgtgga caagagcagg tggcagcagg ggaacgtett ctcatgctcc gtgatgcatg 1380aggctctgca caaccactac acgcagaaga gcctctccct gtctccgggt aaataatcta 1440gagca                                                             1445SEQ ID NO: 2Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly1               5                   10                  15Val His Ser Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys            20                  25                  30Pro Gly Ala Ser Val Lys Val Ser Cys Lys Gly Ser Gly Tyr Thr Phe        35                  40                  45Thr Ser Tyr Trp Met His Trp Val Arg Gln Ala Pro Gly Gln Arg Leu    50                  55                  60Glu Trp Ile Gly Glu Ile Asp Pro Ser Glu Ser Asn Thr Asn Tyr Asn65                  70                  75                  80Gln Lys Phe Lys Gly Arg Val Thr Leu Thr Val Asp Ile Ser Ala Ser                85                  90                  95Thr Ala Tyr Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val            100                 105                 110Tyr Tyr Cys Ala Arg Gly Gly Tyr Asp Gly Trp Asp Tyr Ala Ile Asp        115                 120                 125Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys    130                 135                 140Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly145                 150                 155                 160Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro                165                 170                 175Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr            180                 185                 190Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val        195                 200                 205Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn    210                 215                 220Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro225                 230                 235                 240Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu                245                 250                 255Leu Ala Gly Ala Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp            260                 265                 270Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp        275                 280                 285Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly    290                 295                 300Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn305                 310                 315                 320Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp                325                 330                 335Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro            340                 345                 350Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu        355                 360                 365Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn    370                 375                 380Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile385                 390                 395                 400Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr                405                 410                 415Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys            420                 425                 430Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys        435                 440                 445Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu    450                 455                 460 Ser Leu Ser Pro Gly Lys465                 470 SEQ ID NO: 3gaattctcga gatcgatctc accatgggat ggagctgtat catcctcttc ttggtagcaa   60cagctacagg tgtccactcc gatgtagtga tgactcaaag tccactctcc ctgcctgtca  120cccctggaga accagcttct atctcttgca ggtctagtca gagtcttgca aagagttatg  180ggaacaccta tttgtcttgg tacctgcaga agcctggcca gtctccacag ctcctcatct  240atgggatttc caacagattt tctggggtgc cagacaggtt cagtggcagt ggttcaggga  300cagatttcac actcaagatc tcgcgagtag aggctgagga cgtgggagtg tattactgct  360tacaaggtac acatcagccg tacacgttcg gacaggggac caaggtggag atcaagcgta  420cggtggctgc accatctgtc ttcatcttcc cgccatctga tgagcagttg aaatctggaa  480ctgcctctgt tgtgtgcctg ctgaataact tctatcccag agaggccaaa gtacagtgga  540aggtggataa cgccctccaa tcgggtaact cccaggagag tgtcacagag caggacagca  600aggacagcac ctacagcctc agcagcaccc tgaccctgag caaagcagac tacgagaaac  660acaaagtcta cgcctgcgaa gtcacccatc agggcctgag ctcgcccgtc acaaagagct  720tcaacagggg agagtgttag tctagagcag c                                 750SEQ ID NO: 4Met Gly Trp Ser Cys Ile Ile Leu Phe Leu Val Ala Thr Ala Thr Gly1               5                   10                  15Val His Ser Asp Val Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val            20                  25                  30Thr Pro Gly Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu        35                  40                  45Ala Lys Ser Tyr Gly Asn Thr Tyr Leu Ser Trp Tyr Leu Gln Lys Pro    50                  55                  60Gly Gln Ser Pro Gln Leu Leu Ile Tyr Gly Ile Ser Asn Arg Phe Ser65                  70                  75                  80Gly Val Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr                85                  90                  95Leu Lys Ile Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys            100                 105                 110Leu Gln Gly Thr His Gln Pro Tyr Thr Phe Gly Gln Gly Thr Lys Val        115                 120                 125Glu Ile Lys Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro    130                 135                 140Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu145                 150                 155                 160Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn                165                 170                 175Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser            180                 185                 190Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala        195                 200                 205Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly    210                 215                 220Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys225                 230                 235 SEQ ID NO: 5 Asp Val Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly1               5                   10                  15Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Ala Lys Ser            20                  25                  30Tyr Gly Asn Thr Tyr Leu Ser Trp Tyr Leu Gln Lys Pro Gly Gln Ser        35                  40                  45Pro Gln Leu Leu Ile Tyr Gly Ile Ser Asn Arg Phe Ser Gly Val Pro    50                  55                  60Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile65                  70                  75                  80Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Leu Gln Gly                85                  90                  95Thr His Gln Pro Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys            100                 105                 110Arg Ala Asp Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu        115                 120                 125Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe    130                 135                 140Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln145                 150                 155                 160Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser                165                 170                 175Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu            180                 185                 190Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser        195                 200                 205Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys     210                 215SEQ ID NO: 6Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu1               5                   10                  15Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe            20                  25                  30Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln        35                  40                  45Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser    50                  55                  60Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu65                  70                  75                  80Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser                85                  90                  95Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys            100                 105 SEQ ID NO: 7Arg Ala Asp Ala Ala Pro Thr Val Ser Ile Phe Pro Pro Ser Ser Glu1               5                   10                  15Gln Leu Thr Ser Gly Gly Ala Ser Val Val Cys Phe Leu Asn Asn Phe            20                  25                  30Tyr Pro Lys Asp Ile Asn Val Lys Trp Lys Ile Asp Gly Ser Glu Arg        35                  40                  45Gln Asn Gly Val Leu Asn Ser Trp Thr Asp Gln Asp Ser Lys Asp Ser    50                  55                  60Thr Tyr Ser Met Ser Ser Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu65                  70                  75                  80Arg His Asn Ser Tyr Thr Cys Glu Ala Thr His Lys Thr Ser Thr Ser                85                  90                  95Pro Ile Val Lys Ser Phe Asn Arg Asn Glu Cys            100                 105 SEQ ID NO: 8 Ser Tyr Trp Met His1               5 SEQ ID NO: 9Glu Ile Asp Pro Ser Glu Ser Asn Thr Asn Tyr Asn Gln Lys Phe Lys1               5                   10                  15 GlySEQ ID NO: 10 Gly Gly Tyr Asp Gly Trp Asp Tyr Ala Ile Asp Tyr1               5                   10 SEQ ID NO: 11Arg Ser Ser Gln Ser Leu Ala Lys Ser Tyr Gly Asn Thr Tyr Leu Ser1               5                   10                  15 SEQ ID NO: 12Gly Ile Ser Asn Arg Phe Ser 1               5 SEQ ID NO: 13Leu Gln Gly Thr His Gln Pro Tyr Thr 1               5 SEQ ID NO: 14Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly1               5                   10                  15Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Ser            20                  25                  30Asn Gly Tyr Asn Tyr Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln Ser        35                  40                  45Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro    50                  55                  60Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile65                  70                  75                  80Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Ala                85                  90                  95Leu Gln Thr Pro Gln Thr Phe Gly Gln Gly Lys Val Glu Ile Lys            100                 105                 110 SEQ ID NO: 15 Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala1               5                   10                  15Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr            20                  25                  30Ala Met His Trp Val Arg Gln Ala Pro Gly Gln Arg Leu Glu Trp Met        35                  40                  45Gly Trp Ile Asn Ala Gly Asn Gly Asn Thr Lys Tyr Ser Gln Lys Phe    50                  55                  60Gln Gly Arg Val Thr Ile Thr Arg Asp Thr Ser Ala Ser Thr Ala Tyr65                  70                  75                  80Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys                85                  90                  95Ala Arg Gly Gly Tyr Tyr Gly Ser Gly Ser Asn Tyr Trp Gly Gln Gly            100                 105                 110Thr Leu Val Thr Val Ser Ser         115

1. A method for treating a human subject suffering from primarysclerosing cholangitis (PSC), wherein the method comprises the step of:administering to a subject suffering from PSC, an effective amount of ananti-α4β7 antibody, further wherein the anti-α4β7 antibody has bindingspecificity for the α4β7 complex, wherein the antigen-binding regioncomprises the CDRs: Light chain: CDR1 SEQ ID NO:11, CDR2 SEQ ID NO:12,CDR3 SEQ ID NO:13, Heavy chain: CDR1 SEQ ID NO:8, CDR2 SEQ ID NO:9, andCDR3 SEQ ID NO:10.
 2. The method of claim 1, wherein said effectiveamount is an amount sufficient to normalize serum alkaline phosphatase(ALP) of said subject.
 3. The method of claim 1, wherein said effectiveamount is an amount sufficient to reduce serum alkaline phosphatase(ALP) of said subject by at least 35%.
 4. The method of claim 1, whereinsaid effective amount is an amount sufficient to improve Ishaknecroinflammatory grading score by at least one point.
 5. The method ofclaim 1, wherein the anti-α4β7 antibody is administered to the subjectaccording to the following dosing regimen: a. an initial dose of 300 mgof the anti-α4β7antibody as an intravenous infusion; b. followed by asecond subsequent dose of 300 mg of the anti-α4β7 antibody as anintravenous infusion at about two weeks after the initial dose; c.followed by a third subsequent dose of 300 mg of the anti-α4β7 antibodyas an intravenous infusion at about six weeks after the initial dose; d.followed by a fourth and subsequent doses of 300 mg of the anti-α4β7antibody as an intravenous infusion every four weeks or every eightweeks after the third subsequent dose of the anti-α4β7 antibody asneeded.
 6. The method of claim 1, wherein said subject has chroniccholestatic liver disease with a subsequent diagnosis of PSC.
 7. Themethod of claim 6, wherein the subsequent diagnosis of PSC is based oncholangiographic findings of intrahepatic and/or extrahepatic bile ductirregularities consistent with PSC.
 8. The method of claim 1, whereinsaid subject further has diagnosis of IBD.
 9. The method of claim 8,wherein the diagnosis of IBD is based on clinical and endoscopicevidence and corroborated by a histopathology report.
 10. The method ofclaim 1, wherein said subject does not have a diagnosis of IBD.
 11. Themethod of claim 1, wherein said subject has alkaline phosphatase (ALP)elevation to at least 1.6 times the upper limit of normal (ULN) at thetime of initial treatment.
 12. The method of claim 1, wherein saidsubject's Ishak fibrosis staging score is improved, maintained ornormalized.
 13. The method of claim 1, wherein said subject's AmsterdamCholestatic Complaints (ACCS) has improved, maintained or normalized,5-D itch scale has improved, maintained or normalized, or said subjecthas a liver stiffness TE score of less than or equal to 14.3 kPa, asassessed by Transient Elastrography.
 14. The method of claim 1, whereinsaid anti-α4β7 antibody is administered to said subject intravenously orsubcutaneously.
 15. The method of claim 1, wherein a PSC-related outcomeselected from the group consisting of progression to cirrhosis, liverfailure, death and liver transplantation is delayed or prevented. 16.The method of claim 1, wherein a PSC-related complication selected fromthe group consisting of ascites, hepatic encephalopathy, development ofvarices, jaundice, variceal bleeding, cholangiocarcinoma, hepatocellularcarcinoma, evidence of cirrhosis, and colorectal cancer is delayed orprevented.
 17. The method of claim 1, wherein said treatment does notcause one or more than one adverse event, wherein an adverse event isselected from the group consisting of, hepatoxicity, PML,cholangiocarcinoma, one or more complications due to portalhypertension, leucopenia, lymphopenia, colorectal cancer,infusion-related reactions, infection, acute respiratory failure, acuterespiratory distress syndrome, Torsade de pointer, ventricularfibrillation, ventricular tachycardia, malignant hypertension,convulsive seizure, agranulocytosis, aplastic anemia, toxic epidermalnecrolysis, Stevens-Johnson syndrome, hepatic necrosis, acute liverfailure, anaphylactic shock, acute renal failure, pulmonaryhypertension, pulmonary fibrosis, confirmed or suspected endotoxinshock, confirmed or suspected transmission of infectious agent by amedicinal product, neuroleptic malignant syndrome, malignanthyperthermia, spontaneous abortion, stillbirth, and fetal death.
 18. Themethod of claim 1, wherein said anti-α4β7 antibody is humanized.
 19. Themethod of claim 1, wherein said anti-α4β7 antibody is vedolizumab.20-31. (canceled)